WO2011159178A1 - Nouvelle souche de lactobacillus plantarum s, utilisation de la souche de lactobacillus plantarum s et préparation pour l'ensilage de fourrages grossiers - Google Patents

Nouvelle souche de lactobacillus plantarum s, utilisation de la souche de lactobacillus plantarum s et préparation pour l'ensilage de fourrages grossiers Download PDF

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Publication number
WO2011159178A1
WO2011159178A1 PCT/PL2011/000059 PL2011000059W WO2011159178A1 WO 2011159178 A1 WO2011159178 A1 WO 2011159178A1 PL 2011000059 W PL2011000059 W PL 2011000059W WO 2011159178 A1 WO2011159178 A1 WO 2011159178A1
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Prior art keywords
lactobacillus plantarum
roughages
ensiling
preparation
kkp
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PCT/PL2011/000059
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English (en)
Inventor
Krystyna Zielinska
Krystyna Stecka
Antoni Miecznikowski
Agata Kapturowska
Marta Kuprys
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Instytut Biotechnologii Przemyslu Rolno-Spozywczego
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Application filed by Instytut Biotechnologii Przemyslu Rolno-Spozywczego filed Critical Instytut Biotechnologii Przemyslu Rolno-Spozywczego
Priority to EP11731146.4A priority Critical patent/EP2582851B8/fr
Publication of WO2011159178A1 publication Critical patent/WO2011159178A1/fr
Priority to US13/684,983 priority patent/US8697423B2/en

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C12N1/205—Bacterial isolates
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K10/00—Animal feeding-stuffs
    • A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
    • A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
    • A23K10/18—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K10/00—Animal feeding-stuffs
    • A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K30/00—Processes specially adapted for preservation of materials in order to produce animal feeding-stuffs
    • A23K30/10—Processes specially adapted for preservation of materials in order to produce animal feeding-stuffs of green fodder
    • A23K30/15—Processes specially adapted for preservation of materials in order to produce animal feeding-stuffs of green fodder using chemicals or microorganisms for ensilaging
    • A23K30/18—Processes specially adapted for preservation of materials in order to produce animal feeding-stuffs of green fodder using chemicals or microorganisms for ensilaging using microorganisms or enzymes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/225—Lactobacillus
    • C12R2001/25—Lactobacillus plantarum

Definitions

  • the subject of the invention is a new strain of Lactobacillus plantarum S bacteria, its use and the preparation for ensiling of contaminated roughages.
  • Ochratoxin A is synthesized by moulds from Aspergillus sp. and Penicillium sp., which are often developed on agricultural products. OTA has been classified by the International Agency for Research on Cancer as carcinogenic factor of category 2B. The monitoring studies, as conducted in different years and concerning OTA presence in feedstuffs and food, indicate the existence of serious hazards for animals and humans, especially during hot and wet years, being favorable for growth of moulds.
  • the purpose of the invention was to select a strain of lactic acid bacteria, characterized by the ability of growing in the environment contaminated with ochratoxin A and reducing OTA level by at least 80% in order to apply it in ensiling of the contaminated roughages.
  • the strain according to the invention was isolated from the spontaneous fermentating maize plants. It was classified to Lactobacillus plantarum species on the basis of studying morphological and physiological-biochemical properties with API 50 CH kit (Biomerieux, France) and selected in order to obtain the ability of developing and synthesizing organic acids in growth media, containing ochratoxin A in concentration of 50 - 100 ppb.
  • the object of the invention is the new bacterial strain, which was specified as Lactobacillus plantarum S and deposited in the IAFB Collection of Industrial Microorganisms Institute of Agricultural and Food Biotechnology in Warsaw under the number KKP 2021 p or variant thereof.
  • the second object of the invention is the composition comprising an effective amount of Lactobacillus plantarum S deposited under the number KKP 2021 p or variant thereof.
  • Preferred composition further comprises at least one carrier and/or at least one emulsifier and/or at least bio-stimulator, preferably is in dry form
  • the third object of the invention includes granular or powder preparation for ensiling of the roughages which contains dried-on-the carrier biomass of Lactobacillus plantarum S KKP 2021 p with bacterial number equal to 9 - 10 log CFU/g of the preparation; the composition of the carriers contains extract from aloes which constitutes 0.2 - 2.0 % of dry matter of the preparation.
  • Another object of the invention is a method of ensiling of roughages, which comprises the step of ensiling of roughages with use of new strain of Lactobacillus plantarum S KKP 2021 p according to the invention and/or the composition according to the invention and/or preparation according to the invention.
  • Another object of the invention is the use of the new strain of Lactobacillus plantarum S KKP 2021 p or variant thereof for ensiling of roughages, intended for decontamination of roughages, contaminated with ochratoxin A and/or moulds and/or with pathogenic bacteria selected from genus Salmonella and/or genus Listeria and/or coliform bacteria and/or bacteria of the species Escherichia coli and/or Clostridium perfringens.
  • Another object of the invention is the use of new strain of Lactobacillus plantarum S KKP 2021 p according to the invention and/or the composition according to the invention and/or preparation according to the invention for ensiling of roughages.
  • ensiling of roughages is carried out to decontaminate or/and restrain from contamination and/or prevent contamination of roughages, contaminated with ochratoxin A and/or moulds and/or with pathogenic bacteria selected from genus Salmonella and/or genus Listeria and/or coliform bacteria and/or bacteria of the species Escherichia coli and/or Clostridium perfringens.
  • mutant strain or strains obtained by rising the deposited strain as starter material wherein the mutant strain(s) retain or further improve the ability of growing and activity of parent strain i.e. ability to grow in the environment contaminated with ochratoxin A and reducing OTA level and/or reducing moulds and/or reducing pathogenic bacteria growth.
  • Fig.l Shows changes in ochratoxin A content in medium during cultivation of Lactobacillus plantarum S KKP 2021 p.
  • Fig.2. Shows survival of pathogenic bacteria during their incubation with the strain Lactobacillus plantarum S KKP 2021 p.
  • the new strain according to the invention was isolated from the spontaneous fermentating maize plants. It was classified to Lactobacillus plantarum and deposited in the International Collection of Industrial Microorganism Cultures of the Institute of Agricultural and Food Biotechnology under the number KKP 2021 p.
  • Lactobacillus plantarum S KKP 2021 p strain is characterized by the following features:
  • Bacterial cells have a form of short rods occurring singly, in pairs or in chains of 0.9 - 1.2 um. They do not produce spores and are Gram+.
  • the new strain is relatively heterofermentative, microaerophilic, does not produce catalase, does not reduce nitrates to nitrites, and is capable of fermenting the following sugars: saccharose, galactose, glucose, fructose, arabinose, maltose, mannose, esculin, lactose, cellobiose, gentiobiose, mellibiose, rafinose, ribose, xylose, sorbitol and trehalose.
  • the sequence of 16S rDNA segment of Lactobacillus plantarum S KKP 2021 p is given on SEQ ID NO: l .
  • the new strain is characterized by the ability of developing and synthesizing lactic acid in medium, containing ochratoxin A and of removing it from the environment in at least 80% as well as inhibiting growth of moulds. Moreover, it was revealed that the new strain showed, simultaneously, a very high antibacterial activity against pathogenic bacteria from Salmonella and Listeria genus, Escherichia coli and Clostridium perfringens, which are often present in agricultural products, cultivated on organically fertilized land, especially with the use of liquid organic manure.
  • Antibacterial activity of the strain Lactobacillus plantarum S KKP 2021 p in relation to pathogenic strains Salmonella sp. and Escherichia coli, isolated from digestive tract of sick animals was determined by Pilet method (Pilet et al., 1995, J. Food Prot., 58, 256 - 262). After 24 hours of cultivation of the examined strain, 10 ⁇ of supernatant were introduced onto the surface of agar medium with the diameter of 10 cm, containing Salmonella sp. or Escherichia coli. After the successive 24 hours, the diameter of bacterial growth inhibition zone was measured. In the experimental conditions metabolites, produced by the strain Lactobacillus plantarum S KKP 2021 p, inhibited the growth of tested pathogenic bacteria in the range of 90 - 100%.
  • Bacterial cultures cultivated for 24 h were employed as the experimental material.
  • the cultures of Salmonella sp. and Escherichia coli, with the number of bacteria 6.5 and 6.0 log CFU/ml, respectively, were combined with the culture of Lactobacillus plantarum S KKP 2021 p bacteria with the number of bacteria 9.0 log CFU/ml in v/v ratio 1 :1.
  • the new strain Lactobacillus plantarum S KKP 2021 p inhibited completely Salmonella sp. growth and after seven days of incubation - development of Escherichia coli (see Fig.2).
  • Cultivation of biomass of Lactobacillus plantarum S KKP 2021 p was conducted in fermentors with volume of 150 1, at temperature of 30°C, at pH 5.7, regulated by ammonia water.
  • the content of the components of culture medium in g/1 water was as follows: glucose - 20, yeast extract - 10, maize soak - 5, ammonium biphosphate - 0.2, ammonium sulphate - 0.35 and manganese sulphate - 0.01.
  • Bacteria were cultivated for 24 h. Bacteria biomass with ca.
  • 30% DM obtained after separation of culture medium at rotations of separator equal to 14000/min., was mixed with the following carriers: mix of starter sugars: saccharose, glucose, lactose, soluble potato starch, emulsifier (lecithin) and bio-stimulator (aloes extract). Then, it was dried in fluidal drier at temperature up to 40°C.
  • the dried bacterial preparation contained ca. 94% DM, including: 90% sugars; 0.2% of dried aloes extract and 3.8% of dried bacterial biomass and was characterized by the number of bacteria equal to 9.0 log CFU/g (colonies- forming units).
  • silages In the experimental agricultural farm, two types of silages were produced: from meadow sward and maize plants with humidity of ca. 40%, contaminated with ochratoxin A, moulds and pathogenic bacteria.
  • the preparation, according to the invention obtained in example I
  • the preparation, according to the invention was applied in a form of water spray.
  • the samples of silages without and with the addition of the preparation were collected.
  • the influence of Lactobacillus plantarum S KKP 2021 p on reduction of ochratoxin A, number of moulds and pathogenic bacteria in the ensiled feedstuffs were examined.
  • the number of moulds was lowered by 77% in the silages produced from meadow sward and by 80 % in the silages from maize plants as compared to the silages obtained without the addition of the preparation Lactobacillus plantarum S KKP 2021 p.
  • the contaminated with ochratoxin A and pathogenic microorganisms feeds after the process of lactic fermentation provided by bacteria of the new strain Lactobacillus plantarum S KKP 2021 p, may be employed as completely safe products in feeding of breeding animals.

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  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Health & Medical Sciences (AREA)
  • Microbiology (AREA)
  • Zoology (AREA)
  • Polymers & Plastics (AREA)
  • Biotechnology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Genetics & Genomics (AREA)
  • Organic Chemistry (AREA)
  • Wood Science & Technology (AREA)
  • Animal Husbandry (AREA)
  • Food Science & Technology (AREA)
  • Biomedical Technology (AREA)
  • Biochemistry (AREA)
  • Physiology (AREA)
  • Molecular Biology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Medicinal Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Engineering & Computer Science (AREA)
  • Virology (AREA)
  • Mycology (AREA)
  • Botany (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Fodder In General (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)

Abstract

L'invention concerne la nouvelle souche bactérienne non génétiquement modifiée de Lactobacillus plantarum S, qui a été déposée dans la collection IAFB de l'Industrial Microorganisms Institute of Agricultural and Food Biotechnology de Varsovie sous le numéro KKP 2021 p ou l'un de ses variants. L'invention concerne l'utilisation de la souche de Lactobacillus plantarum S KKP 2021 p ou d'un de ses variants ou d'une composition la comprenant pour l'ensilage de fourrages grossiers, dans l'intention de décontaminer les aliments, contaminés par l'ochratoxine A, des moisissures et des bactéries pathogènes. Les aliments contaminés par des moisissures, l'ochratoxine A et des microorganismes pathogènes, après le procédé de fermentation lactique avec la participation de la bactérie de la nouvelle souche de Lactobacillus plantarum S, peuvent être employés dans la nutrition d'animaux reproducteurs, car ce sont des produits totalement sûrs.
PCT/PL2011/000059 2010-06-16 2011-06-14 Nouvelle souche de lactobacillus plantarum s, utilisation de la souche de lactobacillus plantarum s et préparation pour l'ensilage de fourrages grossiers WO2011159178A1 (fr)

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Application Number Priority Date Filing Date Title
EP11731146.4A EP2582851B8 (fr) 2010-06-16 2011-06-14 Nouvelle souche de lactobacillus plantarum s, utilisation de la souche de lactobacillus plantarum s et préparation pour l'ensilage de fourrages grossiers
US13/684,983 US8697423B2 (en) 2010-06-16 2012-11-26 Strain of Lactobacillus plantarum S, the use of the strain of Lactobacillus plantarum S and the preparation for roughages ensiling

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PLPL391534 2010-06-16
PL391534A PL214583B1 (pl) 2010-06-16 2010-06-16 Szczep bakterii Lactobacillus plantarum S, zastosowanie szczepu bakterii Lactobacillus plantarum S oraz preparat do kiszenia pasz objetosciowych

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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20140178528A1 (en) * 2012-11-22 2014-06-26 Instytut Biotechnologii Przemyslu Rolno-Spozywczego Strain of lactobacillus buchneri a, composition, a multi-component preparation for starch-rich plant preservation, their use and a method for plant preservation
WO2015014372A1 (fr) * 2013-07-30 2015-02-05 OÜ Tervisliku Piima Biotehnoloogiate Arenduskeskus Souche isolée de micro-organisme lactobacillus plantarum tak 59 ncimb42150 et son utilisation
CN105861399A (zh) * 2016-06-17 2016-08-17 浙江大学 预防鸡坏死性肠炎的植物乳杆菌及其应用
CN112143670A (zh) * 2020-09-08 2020-12-29 中国农业科学院农产品加工研究所 一种短乳杆菌8-2b及其应用

Families Citing this family (4)

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KR101470995B1 (ko) * 2013-08-30 2014-12-09 주식회사 바이오리쏘스 프로피온산 생산능을 갖는 미생물 및 그를 포함하는 조사료 조성물
PL241510B1 (pl) 2015-11-06 2022-10-17 Probios Spolka Z Ograniczona Odpowiedzialnoscia Nowy szczep Lactobacillus plantarum AMT14 i kompozycja zawierająca szczep Lactobacillus plantarum AMT14
US10166262B2 (en) 2017-04-20 2019-01-01 Ewelina Sosnowska-Turek Strain of bacteria and composition comprising the same
CN109182220A (zh) * 2018-10-22 2019-01-11 东北农业大学 一株抑制产气荚膜梭菌增殖生长的植物乳杆菌及其应用

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Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20140178528A1 (en) * 2012-11-22 2014-06-26 Instytut Biotechnologii Przemyslu Rolno-Spozywczego Strain of lactobacillus buchneri a, composition, a multi-component preparation for starch-rich plant preservation, their use and a method for plant preservation
US9370199B2 (en) * 2012-11-22 2016-06-21 Instytut Biotechnologii Przemyslu Rolno-Spozywczego Strain of Lactobacillus buchneri A, composition, a multi-component preparation for starch-rich plant preservation, their use and a method for plant preservation
WO2015014372A1 (fr) * 2013-07-30 2015-02-05 OÜ Tervisliku Piima Biotehnoloogiate Arenduskeskus Souche isolée de micro-organisme lactobacillus plantarum tak 59 ncimb42150 et son utilisation
JP2016526900A (ja) * 2013-07-30 2016-09-08 オウ テルヴィスリク ピーマ バイオテウノロジーテ アレンダスケスクス 単離された微生物株ラクトバチルス・プランタルムｔａｋ５９ ｎｃｉｍｂ４２１５０およびその使用
CN105861399A (zh) * 2016-06-17 2016-08-17 浙江大学 预防鸡坏死性肠炎的植物乳杆菌及其应用
CN105861399B (zh) * 2016-06-17 2019-11-05 浙江大学 预防鸡坏死性肠炎的植物乳杆菌及其应用
CN112143670A (zh) * 2020-09-08 2020-12-29 中国农业科学院农产品加工研究所 一种短乳杆菌8-2b及其应用
CN112143670B (zh) * 2020-09-08 2022-07-19 中国农业科学院农产品加工研究所 一种短乳杆菌8-2b及其应用

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Publication number Publication date
EP2582851A1 (fr) 2013-04-24
US20130084623A1 (en) 2013-04-04
EP2582851B8 (fr) 2015-01-07
PL391534A1 (pl) 2011-12-19
US8697423B2 (en) 2014-04-15
EP2582851B1 (fr) 2014-11-19
PL214583B1 (pl) 2013-08-30

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